Methods of inducing immune tolerance with modified anti-cd154 antibodies

Abstract

This disclosure relates to methods of inducing tolerance during transplantation by administering to a transplant recipient anti-CD154 antibodies with modified (e.g., selectively reduced) effector functions, transplanting into the recipient hematopoietic stem cells that produce immune cells that are tolerant of a donor organ, donor tissue or donor cell. The tolerance may be central tolerance, peripheral tolerance, or organ-specific tolerance.

What is claimed: 
     
         1 . A method of inducing immune tolerance in a transplant recipient, the method comprising administering to the recipient one or more doses of an isolated anti-CD154 antibody, transplanting into the recipient hematopoietic stem cells, and transplanting a donor organ, a donor tissue or donor cell into the recipient, wherein the hematopoietic stem cells produce immune cells that are tolerant of the donor organ, donor tissue or donor cell, thereby inducing immune tolerance in the recipient. 
     
     
         2 . The method of  claim 1 , wherein the tolerance is central tolerance. 
     
     
         3 . The method of  claim 2 , wherein the method further comprises depleting T cells in the thymus and/or bone marrow. 
     
     
         4 . The method of  claim 1 , wherein the tolerance is peripheral tolerance. 
     
     
         5 . The method of  claim 4 , wherein the method further comprises depleting T cells in the lymph nodes and/or peripheral tissue. 
     
     
         6 . The method of  claim 1 , wherein the tolerance is organ-specific tolerance. 
     
     
         7 . The method of  claim 6 , wherein the method further comprises depleting T cells from the heart, kidney, liver, lung, pancreas, intestines, islet cells, face, hand, arm, foot, leg or skin or a combination thereof. 
     
     
         8 . The method of any one of  claims 3 ,  5  and  7 , wherein the T cells are depleted by a method selected from a group consisting of total body irradiation, administration of abatacept, administration of one or more BCL-2 inhibitors, administration of busulfan, administration of fludarabine phosphate, administration of cyclophosphamide, administration of one or more immunosuppressive T cell-depleting antibodies, administration of one or more anti-αβ T cell receptor antibodies, and administration of one or more CD122 antagonists or a combination thereof. 
     
     
         9 . The method of  claim 8 , wherein the one or more T cell-depleting antibodies are selected from the group consisting of anti-CD4, anti-CD8, anti-CD45, anti-CTLA4, anti-CD20, and anti-CD33 antibodies or a combination thereof. 
     
     
         10 . The method of any one of  claims 1 - 9 , wherein the method results in mixed chimerism. 
     
     
         11 . The method of any one of  claims 1 - 10 , wherein the anti-CD154 antibody is administered prior to, subsequently to or simultaneously with the transplantation of the hematopoietic stem cells. 
     
     
         12 . The method of  claim 1 - 11 , wherein the anti-CD154 antibody is administered at a dose of 5-50 mg/kg. 
     
     
         13 . The method of any one of  claims 1 - 12 , wherein the anti-CD154 antibody is administered subcutaneously, intravenously, intravitreally, orally, via inhalation, transdermally, or rectally. 
     
     
         14 . The method of any one of  claims 1 - 13 , wherein the anti-CD154 antibody, comprises a human or humanized variable domain, wherein the variable domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), and wherein the VH is operably linked to a human Fc domain with modified effector functions. 
     
     
         15 . The method of  claim 14 , wherein one or more effector functions are reduced. 
     
     
         16 . The method of  claim 14  or  15 , wherein one or more effector functions are eliminated. 
     
     
         17 . The method of any one of  claims 14 - 16 , wherein the VH is operably linked to a human Fc region, wherein the human Fc region comprises a human hinge sequence and the human Fc domain, wherein the human hinge sequence is between the VH and the human Fc domain. 
     
     
         18 . The method of  claim 17 , wherein the hinge comprises the amino acid sequence of any one of SEQ ID NOs: 76-90. 
     
     
         19 . The method of any one of  claims 14 - 18 , wherein the Fc domain is derived from an IgG4 Fc (or crystallizable fragment) region. 
     
     
         20 . The method of  claim 19 , wherein the Fc domain comprises one or more amino acid modifications that modifies effector functions. 
     
     
         21 . The method of  claim 20 , wherein the antibody comprises an amino acid modification at any one of the positions selected from the group consisting of S228, L235, G237, E318, and N297 or a combination thereof, wherein the numbering of amino acid residues is according to the EU index as set forth in Edelman. 
     
     
         22 . The method of  claim 21 , wherein the antibody comprises an amino acid modification selected from the group consisting of S228P, F234A, L235A, L235E, G237A, E318A, and N297Q or a combination thereof. 
     
     
         23 . The method of any one of  claims 14 - 18 , wherein the Fe domain is derived from an IgG1 Fe (or crystallizable fragment) region and comprises one or more amino acid modifications that modifies effector functions. 
     
     
         24 . The method of  claim 23 , wherein the antibody comprises an amino acid modification at any one of the positions selected from the group consisting of E216, R217, K218, C219, C220, C226, C229, P230, E233, L234, L235, G236, G237, P238, S239, V240, F241, K246, L251, T260, D265, V266, H268, W277, N297, E318, K322, P329, A330, P331, Q347, N348, T350, L351, K360, T366, N390, K392, T394, D399, S400, F405, Y407, K409, T411, or a combination thereof, wherein the numbering of amino acid residues is according to the EU index as set forth in Edelman. 
     
     
         25 . The method of  claim 24 , wherein the antibody comprises an amino acid modification selected from the group consisting of C220S, C226S, C229S, P230S, E233P, L234A, L234F, L234V, L235A, L235E, L235V, G236E, G237A, P238S, D265S, D265A, H268Q, W277T, N297G, N297Q, N297D, N297A, E318A, K322A, P329G, P329A, A330S, P331S, Q347R, Q347E, Q347K, T350V, L351Y, K360D, K360E, T366A, T366I, T366L, T366M, T366V, N390R, N390K, N390D, K392V, K392M, K392R, K392L, K392F, K392E, T394W, D399R, D399W, D399K, S400E, S400D, S400R, S400K, F405A, F405I, F405M, F405T, F405S, F405V, F405W, Y407A, Y407I, Y407L, Y407V, K409F, K409I, K409S, K409W, T411N, T411R, T411Q, T411K, T411D, T411E, T411W, AE216-E222, K246R/L251E/T260R, InR234/235, InV235/236, InR236/237, InR237/238, InV238/239, InN238/239, InL238/239, InE238/239, InG238/239, InS239/240, InG240/241, InE240/241, InG240/241, InL238/239/P238Q, InE238/239/N348A, InS239/240/V266A, and InR237/238/G236A or a combination thereof. 
     
     
         26 . The method of any one of  claims 14 - 18 , wherein the Fc domain is derived from an IgG2 Fc (or crystallizable fragment) crystallizable fragment region. 
     
     
         27 . The method of  claim 26 , wherein the antibody comprises an amino acid modification at any one of the positions selected from the group consisting of V234, G237, P238, H268, V309, A330, and P331 or a combination thereof, wherein the numbering of amino acid residues is according to the EU index as set forth in Edelman. 
     
     
         28 . The method of  claim 27 , wherein the antibody comprises an amino acid modification selected from the group consisting of V234A, G237A, P238S, H268Q, H268A, V309L, A330S, and P331S, or a combination thereof. 
     
     
         29 . The method of any one of  claims 14 - 18 , wherein the Fe domain comprises the amino acid sequence selected from the group consisting of SEQ ID NOs: 3-9, 12-18, and 238-241. 
     
     
         30 . The method according to  claim 17 , wherein the Fc region comprises the amino acid sequence selected from the group consisting of SEQ ID NOs: 21-37, 40-56, and 243-251. 
     
     
         31 . The method of any one of  claims 14 - 30 , wherein the VH comprises:
 (a) a heavy chain CDR1 having the amino acid sequence of SEQ ID NO: 57,   (b) a heavy chain CDR2 having the amino acid sequence of SEQ ID NO: 58, and   (c) a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 59; and   
       wherein the VL comprises:
 (a) a light chain CDR1 having the amino acid sequence of SEQ ID NO: 60, 
 (b) a light chain CDR2 having the amino acid sequence of SEQ ID NO: 61, and 
 (c) a light chain CDR3 having the amino acid sequence of SEQ ID NO: 62. 
 
     
     
         32 . The method of any one of  claims 14 - 31 , wherein the VH comprises the amino acid sequence selected from the group consisting of SEQ ID NO: 63, 64, 252 and 253. 
     
     
         33 . The antibody of any one of  claims 14 - 32 , wherein the VL comprises the amino acid sequence of SEQ ID NO: 65 or 66. 
     
     
         34 . The method of any one of  claims 17 - 33 , wherein the antibody further comprises a CH1 domain, wherein the CH1 domain is operably linked to
 (a) the C-terminal end of the VH, and   (b) the N-terminal end of the hinge.   
     
     
         35 . The method of  claim 34 , wherein the CH1 domain comprises an amino acid sequence that is at least 80% identical to the amino acid sequence of any one of SEQ ID NOs: 67, 70, and 73. 
     
     
         36 . The method of any one of  claims 14 - 35 , wherein the antibody comprises a linker between the VH and the Fc domain. 
     
     
         37 . The method of any one of  claims 17 - 35 , wherein the antibody comprises a linker between the VH and the hinge. 
     
     
         38 . The method of  claim 34  or  35 , wherein the antibody comprises a linker between the VH and the CH1 domain. 
     
     
         39 . The method of any one of  claims 36 - 38 , wherein the linker comprises the amino acid sequence of any one of SEQ ID NOs: 199-223 and 327-330. 
     
     
         40 . The method of any one of  claims 14 - 39 , wherein the VH is operably linked to the amino acid sequence of any one of SEQ ID NOs: 3-9, 12-18, 21-37, 40-56, 238-241, and 243-251. 
     
     
         41 . The method of any one of  claims 14 - 40 , wherein the heavy chain comprises the amino acid sequence of any one of SEQ ID NOs: 121-132, 135-146, 149, 151-160, 163, 165-174, 266-277, and 279-288. 
     
     
         42 . The method of any one of  claims 14 - 41 , wherein the light chain comprises the amino acid sequence of SEQ ID NO: 195 or 196. 
     
     
         43 . The method of any one of  claims 14 - 42 , wherein the antibody is monoclonal. 
     
     
         44 . The method of any one of  claims 14 - 43 , wherein the antibody is chimeric. 
     
     
         45 . The method of any one of  claims 14 - 44 , wherein the antibody is humanized. 
     
     
         46 . The method of any one of  claims 14 - 30 ,  34 - 40  and  43 , wherein the antibody is human. 
     
     
         47 . The method of any one of  claims 14 - 46 , wherein the binding of the antibody to human CD154 inhibits the interaction between human CD154 and human CD40. 
     
     
         48 . The method of any one of  claims 14 - 47 , wherein the antibody blocks the activation of one or more of B cells, macrophages, dendritic cells, or endothelial cells by inhibiting binding of CD154 to CD40. 
     
     
         49 . The method of any one of  claims 14 - 48 , wherein the antibody has one or more of the following effects when administered to a subject:
 (a) decreased risk of thrombosis or thromboembolic events compared to hu5c8 antibody;   (b) decreased activation of platelets expressing CD154;   (c) inhibition of CD154 shedding; and   (d) alteration of the expression or activity of downstream targets of CD154-CD40 signaling.   
     
     
         50 . The method of any one of  claims 14 - 49 , wherein the antibody has a K D  of less than 50 pM for CD154, such as 5-25 pM or 9.5-23 pM. 
     
     
         51 . The method of any one of  claims 14 - 50 , wherein the antibody does not comprise the amino acid sequence consisting of any one of SEQ ID NOs: 119, 120, 133, 134, 147, 148, 150, 161, 162, 164, 230, 234 and 278. 
     
     
         52 . The method of any one of  claims 1 - 51 , wherein the transplant recipient is human. 
     
     
         53 . The method of any one of  claims 1 - 51 , wherein the transplant recipient is non-human. 
     
     
         54 . The method of  claim 53 , wherein the transplant recipient is a monkey. 
     
     
         55 . The method of any one of  claims 1 - 54 , wherein the transplant is an allogeneic transplant, autologous transplant or a xenogeneic transplant. 
     
     
         56 . The method of any one of  claims 1 - 54 , wherein the donor cell is an engineered cell or an ex-vivo expanded cell. 
     
     
         57 . The method of  claim 56 , wherein one or more genes in the donor cell is modified using one or more techniques selected from a group consisting of transduction to express a cDNA, a CRISPR/Cas9 system, RNAi technology and retroviral technology. 
     
     
         58 . The method of  claim 56  or  57 , wherein the donor cell is modified to express a chimeric antigen receptor (CAR) on its surface. 
     
     
         59 . The method of any one of  claims 55 - 58 , wherein the donor cell is selected from a group consisting of a stem cell, a regulatory T cell, a CAR-T cell, a CAR-B cell, a tumor-infiltrating lymphocyte (TIL). 
     
     
         60 . The method of any one of  claims 1 - 59 , wherein the method promotes a long-term survival of the donor organ, donor tissue or donor cell, wherein said long-term graft survival is selected from the group consisting of:
 (a) at least 6 months post-transplant;   (b) at least 1 year post-transplant; and   (c) at least 5 years post-transplant.   
     
     
         61 . The method of any one of  claims 1 - 60 , wherein the anti-CD154 antibody is administered locally. 
     
     
         62 . The method of any one of  claims 1 - 60 , wherein the anti-CD154 antibody is administered systemically.